I'm going to start to work with this system and I have a question, can I prepare the Ascorbate and store it in the fridge for a while or its better to prepare all the time fresh for the assay? Im using L-acid ascorbic, is it ok?
Always prepare fresh. Ascorbate rapidly oxidises in physiological buffers (exposed to air) so it is best to dissolve it just before you make up the reaction. For convenience you can aliquot some pre-weighed amounts of the powder and store them in sealed tubes ready to be dissolved (try to keep them in a vacuum bell or with some dessicant). Ascorbate is simply the anion of ascorbic acid that predominates at neutral pH (the acid form becomes the ascorbate form in physiological buffers at neutral pH)
If you dissolve ascorbic acid in 1 mM aqueous HCl it will last a bit longer as this will slow down oxidation by air. Still, I'd never use ascorbate solution prepared the previous day. And if you flush the solid ascorbate with nitrogen or argon gas it will keep a long time.
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